This kit provides a fast, reliable and convenient method to purify and concentrate high quality, high purity and inhibitor-free cell-free circulating DNA, circulating including exosomal RNA as well as viral DNA/RNA from fresh, frozen or preserved urine samples from volumes ranging from 2 mL to 10 mL.

Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix. The kit is designed to isolate all sizes of cfc-DNA and circulating RNA, including microRNA, as well as all sizes of exosomal RNA. Norgen’s Urine Cell-Free Circulating and Viral Nucleic Acid Purification Kits provides a clear advantage over other available kits in that they do not require phenol/chloroform or any protease treatments. Moreover, the kit allows the user to elute into a flexible elution volume ranging from 50 µL to 100 µL. The purified nucleic Acid is of the highest integrity, and can be used in a any downstream applications including PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, methylation-sensitive PCR and Southern Blot analysis, Northern blotting, RNase protection and primer extension, expression array assays, and NGS.

Background

Recent evidence indicates that cell-free circulating (cfc) nucleic acids such as DNA and RNA, including exosomal RNA, in urine contains valuable information for the discovery of biomarkers that can help with early detection of certain cancer types, for monitoring the disease status as well as for the detection fetal DNA. Moreover, urine has been widely used for the detection of viral infections, including HIV, HCV and HBV.The advantage for using urine as a source for cancer biomarkers is that it can be acquired in large quantities without using invasive procedures, and repeated sampling from the same individual is applicable, which facilitates longitudinal studies. There are many advantages favouring the use of urinary nucleic acids for cancer biomarker discovery over blood, tissue samples or other bodily fluids, including: (1) urine is non-infectious for HIV and less infectious for many other pathogens; (2) the profile of urinary nucleic acids is similar to that in urine or serum; (3) nucleic acid purification from urine is technically much easier because of its low protein concentration (1000-fold lower than blood).

Kit Specifications

Minimum Urine Input

2 mL

Maximum Urine Input

10 mL

Size of Nucleic Acid Purified

All sizes, including miRNA and
small RNA (< 200 nt)

Elution Volume

50-100 μL

Time to Complete 10 Purifications

40-45 minutes

Average Yields

Variable depending on specimen

Storage Conditions and Product Stability

All buffers should be kept tightly sealed and stored at room temperature (15-25°C) for up to 2 year without showing any reduction in performance. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.